Part:BBa_K3520010

GFP Superfolder for Flavobacteriia

iGEM KU Istanbul 2020

This part was designed during the Partnership of iGEM Athens 2020 and iGEM KU Istanbul 2020. The latter team is creating a communication scheme between humans and biological cells by morphing cells into lasers. By this technology, they will be able to detect changes inside and around cells and tissues. These cell lasers can be employed in diagnostics and therapeutic purposes alongside as a high throughput method in basic research.

iGEM Athens 2020

iGEM Athens 2020 team during the project MORPHÆ works with Flavobacteriia to produce a non-cellular structurally coloured biomaterial which will require the secretion of a biomolecule that Flavobacteriia do not normally secrete. Our hypothesis is that the formed matrix will have a structure similar to that of the biofilm and thus, it will provide the material with macroscopically the same colouration properties as the biofilm.

So these two teams above, collaborated in a creative way and iGEM Athens designed a cloning experiment in which Flavobacteriia will express reflectin with a signal peptide which will translocate it to the outer membrane and GFP superfolde. As a result, the biolaser designed by iGEM KU Istanbul will be able to track genetically modified Flavobacteriia.

Further information about Biolaser's Function

Biological laser or biolaser is an emerging concept with huge potential in biological and biomedical research. Biolaser is a new type of laser which has biological materials as part of its gain medium or optical feedback. Biolasers are very close to fluorescence microscopy where active molecules (fluorescent) emit light upon excitation by an external laser. However, biolasers are different from fluorescence with respect to optical feedback and gain. The light emitted from a biolaser is coherent, meaning it provides a high signal to noise ratio, consists of a narrow spectrum, and can focus on a small spot, whereas fluorescence radiation is dispersive and very weak compared to biolasers. We are developing a biological laser concept in which no artificial cavities will be used to confine light as opposed to conventional biolasers. Cavities we will be using are composed of natural biological materials which cover the membrane of the cell. So the biolaser concept is fully composed of biological materials. This goal will be achieved by genetically modifying cells to express silicatein and reflectin proteins as well as fluorescent proteins. Expressed silicatein and reflectins will be transported toward the membrane of the cell, and these proteins will be covering the inside and/or outside of the cell.

GFP superfolder

GFP superfolder is a variant of green fluorescent protein (GFP). GFPs often misfold when expressed as fusions with other proteins so a robustly folded version of GFP has been generated called 'superfolder' GFP. GFP superfolder folds well even when fused to poorly folded polypeptides. Compared to 'folding reporter' GFP, a folding-enhanced GFP containing the 'cycle-3' mutations and the 'enhanced GFP' mutations F64L and S65T, superfolder GFP shows improved tolerance of circular permutation, greater resistance to chemical denaturants and improved folding kinetics. The fluorescence of Escherichia coli cells expressing each of eighteen proteins from Pyrobaculum aerophilum as fusions with superfolder GFP was proportional to total protein expression. In contrast, fluorescence of folding reporter GFP fusion proteins was strongly correlated with the productive folding yield of the passenger protein. X-ray crystallographic structural analyses helped explain the enhanced folding of superfolder GFP relative to folding reporter GFP.